1. Field of the Invention
The present invention relates to an isolation and purification method of biomolecules using a hydrogel.
2. Description of the Related Art
Isolation methods of DNA from cells were performed using materials that have the proclivity of binding to DNA. Examples of materials for isolation methods of DNA are silica, glass fiber, anion exchange resin and magnetic beads (Rudi, K. et al., Biotechniqures 22, 506-511 (1997); and Deggerdal, A. et al., Biotechniqures 22, 554-557 (199)). To avoid the manual steps and remove operator error, several automatic machines were developed for high-throughput DNA extraction.
Conventionally, a method of purifying nucleic acids using a solid phase was known. For example, U.S. Pat. No. 5,234,809 discloses a method of purifying nucleic acids using a solid phase to which nucleic acids are bound. However, this method is time consuming and complicated, and thus is not suitable for a Lab-On-a-Chip (LOC). The method also has a problem in the use of a chaotropic material. That is, when the chaotropic material is not used, nucleic acids are not bound to the solid phase.
U.S. Pat. No. 6,291,166 discloses a method of archiving nucleic acids using a solid phase matrix. This method is advantageous in that since nucleic acids are irreversibly bound to the solid phase matrix, a delayed analysis or repeated analysis for the nucleic acid solid phase matrix complexes is possible. However, according to this method, aluminum (Al) which has a positively-charged surface should be rendered hydrophilic with basic materials, such as NaOH, and nucleic acids are irreversibly bound to the Al rendered hydrophilic, and thus cannot be separated from Al.
U.S. Patent Publication No. 2001/18513 discloses a method for extracting biomolecules from a biological sample, the method including: at a first pH, bringing the biological sample into contact with a solid phase such that the biomolecules are bound to the solid phase; and extracting the biomolecules bound to the solid phase using an elution solvent at a second pH. This method is a DNA isolation method using a material with the charge varied according to the pH and does not include the isolation of biomolecules using a hydrogel.
U.S. Pat. No. 5,705,628 discloses a method of reversibly and non-specifically binding DNA to magnetic microparticles having a carboxyl group-coated surface, the method including: combining magnetic microparticles having a carboxyl group-coated surface to a solution containing DNA; and adding the salt and polyethylene glycol onto the surfaces of the magnetic microparticles. However, in the method, the magnetic particles having a carboxylic group-coated surface, the salt and polyethylene glycol are used to isolate DNA, but the isolation of biomolecules using a hydrogel is not included.
Conventional isolation and purification methods of biomolecules requires external devices, such as an electromagnet, and had a problem of practicing time of at least about 30 min. Thus, a method of efficiently isolating and purifying biomolecules within 5 min without an external device is required.
Thus, the present inventors discovered that when biomolecules were isolated using a hydrogel which is ionized in a solution to have a varying surface, the isolation time can be shortened and an external device is not required, and thus this method can be easily applied to Microsystems.